Effects of corticoliberin on synaptic transmission in rat olfactory cortex slices in a water immersion model of depression.

Corticoliberin (corticotrophin-releasing factor, CRF, CRH) is an active regulator of endocrine, autonomic, and immune functions in stress, as well as a mediator of anxiety, determining the behavioral stress response. The present report describes studies of its action on neuron activity evoked by microstimulation of olfactory cortex slices. Behavioral testing in a T maze was used to select individuals with a passive behavioral strategy from a population of Wistar rats, and the animals were subjected to water immersion. Olfactory cortex slices were prepared 10 days later and evoked focal potentials were recorded on perfusion with medium containing corticoliberin (0.1 microM). Among active rats, 60% of slices retained high excitability after stress, and corticoliberin produced only insignificant reductions in the amplitudes of excitatory potentials in these slices, simultaneously increasing the amplitudes of inhibitory potentials. Low excitability was found in 40% of slices from active stressed rats, and corticoliberin had a significant inhibitory effect in these slices. Addition of corticoliberin to the incubation medium used for slices from passive rats with initially low excitability led to complete blockade of synaptic transmission. These data support the involvement of corticoliberin in the development of depression.

Effects of exogenous application of corticotropin-releasing hormone to slices of the olfactory cortex from rats with an active strategy of adaptive behavior on the water-immersion model of depression.

Experiments were performed on male Wistar rats. The specimens with an active strategy of behavior were exposed to unavoidable water-immersion stress. Surviving slices of the olfactory cortex were obtained 10 days after stress. The neurohormone had a strong inhibitory effect in 40% slices from active rats. The activity of glutamate receptors decreased, while the function of GABA receptors increased in 60% slices. Our results indicate that the depressive state of behaviorally active animals due to exposure to unavoidable stress is not necessarily mediated by the corticoliberinergic mechanisms in cortical structures.

Free-radical lipid oxidation in the brains of active and passive rats during the development of post-stress depression.

Free-radical lipid oxidation was studied in the cerebral cortex, striatum, hippocampus, and hypothalamus of KHA and KLA rats (Koltushi High and Low Avoidance) during the development of post-stress depression. After unavoidable emotional-pain exposure, changes in the free-radical oxidation of lipids were phasic in nature and had a clear structural specificity in the early phases. During the maximum development of depression, the most marked impairments to lipid peroxidation were seen in KHA rats in the striatum and hippocampus, while the greatest changes in KLA rats were seen in the striatum and hypothalamus. These data support the important role of the initial behavioral strategy in the pathogenetic mechanisms forming post-stress psychopathology.

Electrophysiological characteristics of depressive states in rats with passive strategies of adaptive behavior.

The behavior of rats in a T-maze was used to select individuals with a passive strategy of adaptive behavior from a population of Wistar rats. These animals were subjected to water immersion and olfactory cortex slices were prepared from the brain 10 days later and used for recording of evoked focal potentials and the effects of tetanic stimulation. Postsynaptic potentials, of both the AMPA and NMDA types, were initially of decreased amplitude in passive rats. After unavoidable stress, the suppression of excitatory potentials increased and there was a simultaneous increase in the amplitude of inhibitory GABAB-ergic mechanisms. Addition of corticoliberin (10(-10) M) to the incubation medium led to reversible blockade of synaptic transmission. Tetanic stimulation of slices from stressed rats led to the development of posttetanic depression in 84% of cases and to post-tetanic potentiation in 12%; this is evidence for profound changes in synaptic transmission. Thus, activation of corticoliberinergic mechanisms in cortical structures does not promote recovery from depressive states in rats with the passive behavioral strategy induced by unavoidable stress.

Blockade of corticoliberin receptors prevents the development of post-stress psychopathology in rats with an active strategy of adaptive behavior.

Active and passive Wistar rats were subjected to single water immersions, after which they showed signs of post-stress depression. Administration on this background of the peptide CRH-R1 receptor blocker astressin prevented the development of behavioral deficit in active individuals but had no effect on the behavior of passive rats. These results lead to the conclusion that corticoliberin receptor blockers are effective in the treatment of post-stress depression only for individuals with an initially active behavioral strategy.

Changes in the adaptive behavior of active and passive wistar rats in a water immersion model of depression.

Animals with active and passive strategies of adaptive behavior were selected from a population of Wistar rats by testing in a T maze to measure the indexes of behavioral passivity and behavioral activity. After single (stress) or two (stress-restress) water immersions, individual changes in adaptive behavior were used to study the development of post-stress psychopathology and its interaction with the initial behavioral strategy. In the unavoidable aversive environment, active and passive rats developed different types of post-stress depression, only passive individuals fulfilling the criteria of post-traumatic stress disorder.

Reactivity of the hypophyseal-adrenocortical system to stress in rats with active and passive behavioral strategies.

Rats with active (KHA) and passive (KLA) behavioral strategies showed no strain-related differences in basal corticosterone levels or in changes in corticosterone levels after exposure to mildly stressful stimuli. Only severe immobilization stress produced significant interstrain differences in the reactivity of the hypophyseal-adrenocortical system, as evidenced by the greater increase in blood corticosterone in KHA rats 30 min after stressing. The hormonal stress response in KHA rats was prolonged, as the elevated blood corticosterone level in these animals persisted longer than in KLA rats. The data provide evidence not only that the hypophyseal-adrenocortical systems have different sensitivities to severe stresses in these strains, but also that active and passive animals have different rates of inactivation of the stress reaction.

Adaptive behavior in active and passive rats after intranasal administration of corticotrophin-releasing hormone.

Rats with high (KHA) and low (KLA) rates of acquiring active avoidance reflexes were used to study the effects of intranasal administration of corticotrophin-releasing hormone on orientational-investigative behavior in an open field and anxiety in an elevated cross maze. Administration of the neurohormone induced opposite changes in the behavior of the rats of these lines in the two tests. In KLA rats, movement and investigative activity increased, while in KHA rats these behaviors decreased. In the elevated maze, KLA rats, unlike KHA rats, showed increases in the time spent in the open arms, which was evidence for a decrease in anxiety in these animals. Thus, intranasal hormone administration completely reproduced the effects seen after administration into the striatum. It is suggested that corticotrophin-releasing hormone is an endogenous factor for the detailed and appropriate correction of adaptive behavior.

Neurophysiological effects of corticotropin-releasing factor in living slices of the olfactory area of the rat cortex.

Application of corticotropin-releasing factor (CRF) at concentrations of 10(-9) and 10(-8) M to living brain slices induced activation of the pre- and postsynaptic excitatory components of focal potentials recorded in the slices. The amplitudes and durations of the AMPA and NMDA components of EPSP increased during exposure to CRF, while the amplitude of the GABA(B)-mediated IPSP was suppressed. At the higher CRF concentration (10(-8) M), cells in slices showed epileptiform discharges. The effects of CRF were reversible and disappeared on washing. Long-term treatment with CRF (90 min) induced changes in cells in slices similar to those associated with long-term post-tetanic potentiation. These data provide evidence that CRF has marked activatory properties and influences the glutamatergic and GABAergic systems.

Involvement of dopaminergic processes in the striatum during the effects of corticoliberin on the behavior of active and passive rats.

The effects of intranasal corticoliberin on behavior in the open field test were studied in rats with active and passive behavioral strategies (lines KHA and KLA); levels of dopamine and noradrenaline and their metabolites were measured in the striatum and hypothalamus. In KLA rats, administration of the neurohormone led to increases in motor and investigative activity, while decreases were seen in KHA rats. There were no interline differences in catecholamine levels in the hypothalamus, while dopamine levels in the KLA striatum nearly doubled and metabolite levels (DOPAC, HVA) were significantly lower than in KHA rats. Corticoliberin increased dopamine and noradrenaline levels in the hypothalamus of both rat lines, with significant decreases in the striatum. This decrease was more marked in KLA rats, probably due to the faster metabolism of transmitters in the presence of neurohormones, as indicated by the increase in metabolite levels in this structure.

Corticoliberin protects neurons from the negative influences of "dysfunctins" in living olfactory cortex slices.

The protective effects of corticoliberin on living rat olfactory cortex slices during perfusion with "dysfunctins" extracted from cerebrospinal fluid of drug addicts were studied. Isolated perfusion of slices with medium containing "dysfunctins" led to irreversible suppression of the amplitude of individual components of focal potentials induced by electrical stimulation of the lateral olfactory tract. The maximum level of depression was seen for the AMPA and NMDA components of EPSP. Preliminary perfusion of slices with medium containing corticoliberin (100 nM) for 15 min partially, and for 30 min completely protected processes mediated by activation of AMPA and NMDA receptor mechanisms from the negative influences of "dysfunctins." It is suggested that corticoliberin can induce its protective effects either via its own specific receptors or non-specifically via glutamate receptors. It is also possible that both of these mechanisms act in combination.

Localization of corticoliberin receptors in the rat brain.

In situ hybridization was used to study the distribution of corticoliberin receptors of subtypes 1 and 2 (CL-R1 and CL-R2 respectively) in different structures of the rat brain. Levels of CL-R1 mRNA in the brain were significantly greater than levels of CL-R2 mRNA, and the most intense expression of the CL-R1 gene was seen in forebrain structures, especially various neocortical, archicortical, and paleocortical regions in the cerebellar cortex. In addition, significant levels of CL-R1 mRNA expression were noted in the red nucleus and the reticular nucleus of the tegmentum. Intense expression of CL-R2 mRNA was observed in structures of the olfactory system, corticomedial parts of the amygdala, fields CA1-CA4 of the hippocampus, the ventromedial hypothalamus, and several brain stem nuclei. Moderate levels of CL-R2 mRNA were seen in the dorsolateral neostriatum. These results provide evidence that corticoliberin receptors of both subtypes are widespread in the brain. The different patterns of expression of CL-R1 and CL-R2 in the brain probably provide the basis for the functional specificity of action of corticoliberin in brain structures.

The role of sex steroids in forming anxiety states in female mice.

Natural fluctuations in sex hormones during the ovarian cycle have enormous influences on ongoing psychological status in the female body. We report here studies of the effects of exogenous sex steroids on anxiety levels in female mice, as evaluated in the elevated cross maze test. Female NMRI mice were subjected to bilateral oophorectomy and one week later received s.c. injections of solvent (sesame oil, controls) or estradiol benzoate for 7 days, either alone or with an additional dose of progesterone on day 7. Elevated maze tests performed 6 h later showed that animals given progesterone had the highest levels of anxiety and the highest levels of grooming reactions as compared with the other groups. Immunohistochemical analysis of the distribution of progesterone receptors in different parts of the brain demonstrated significant increases in the numbers of immunopositive cells after injections of estradiol benzoate alone, with further increases after progesterone injections. Thus, the data obtained here suggest that the genomic effects of sex steroids are important, as they appear to be involved in non-sexual forms of behavior, particularly the level of anxiety.

Expression of early genes in the rat brain after administration of corticoliberin into the neostriatum.

In situ hybridization using oligonucleotide probes was used to study the effects of intrastriatal microinjection of corticoliberin on the expression of the early genes c-fos, jun B, c-jun, and NGFIA in the rat brain. Administration of corticoliberin (0.25 microg) into the neostriatum induced the expression of mRNA encoded by the early genes c-fos, jun B, and NGFIA in both the neostriatum itself and in its efferent structures, particularly the nucleus accumbens and various parts of the cortex. Intrastriatal microinjection of corticoliberin had no effect on the expression of mRNA for the oncogene c-jun in the brain. These results suggest that neuronal activation in the neostriatum and its projection targets manifest as the expression of early genes is one of the mechanisms underlying the adaptive effects of corticoliberin in stress.

The neonatal glucocorticoid treatment-produced long-term changes of the pituitary-adrenal function and brain corticosteroid receptors in rats.

Two distinct periods of sensitivity to elevated glucocorticoid hormone levels during postnatal development of the pituitary-adrenal axis were studied. Wistar rats were injected subcutaneously (s.c.) with cortisol (1 mg/kg) on postnatal days 1-5 or 14-18. The steroid treatment during the first postnatal week resulted in a decrease of the morning basal and stress-induced plasma corticosterone levels in 30 day-old male rats, as well as in rats that were injected with cortisol on the third postnatal week. Stress-induced corticosterone levels in 90-day old cortisol-treated rats were determined in blood samples drawn from the tail vein before the restraint stress, immediately after the 20-min long stress, then 60 and 180 min afterwards. Only the rats treated with cortisol during the third week showed a prolonged stress-induced corticosterone secretion, with the highest corticosterone level in 180 min after the restraint stress. The early neonatal cortisol treatment had no effect on (3)H-corticosterone binding in all studied brain areas of the 90-day old rats. The rats treated with cortisol at the 14-17th postnatal days showed a significantly lower (3)H-corticosterone binding in the frontal cortex, hippocampus, and hypothalamus. These findings suggest that the third week of life in rats is more sensitive to elevated levels of corticosterone than the first one. The high level of glucocorticoids at this period has long-term effects on the efficiency of the negative feedback mechanisms provided by hypothalamus-pituitary-adrenal axis.

Involvement of the striatum in the central regulation of the hormonal functions of the gonads.

Studies reported here show that intrastriatal administration of corticoliberin to rats decreases the blood testosterone level. However, in conditions of chemical deficiency of dopaminergic transmission in the dorsal striatum induced by injection of 6-hydroxydopamine, the effect of this neurohormone did not appear. It is concluded that extrahypothalamic corticoliberin is involved in regulating the hormonal reproductive system acting via dopaminergic mechanisms.

Dopaminergic mechanisms of neostriatum in the regulation of adaptive behavior by corticoliberin.

A conditioned active avoidance response was developed in rats with high (KHA) and low (KLA) rates of learning and the effects of injection of corticoliberin into the dorsal striatum on orientational-investigative and avoidance behavior were studied in normal animals and after depletion of striatal dopamine by preliminary injection of 6-hydroxydopamine. These studies showed that corticoliberin, like 6-hydroxydopamine, produced similar trends in the animals' behavior. Their effects were mediated by opposite mechanisms in animals with initial active and passive learning strategies for adaptive behavior. The role of dopaminergic structures of the striatum in mediating the behavioral effects of corticoliberin is discussed.

Striatal mechanism of action of corticoliberin on behavior in dogs in conditions of dopamine deficiency.

This report describes studies of the interaction of the integrative dopaminergic and corticoliberin systems in the neostriatum during performance of situational food-related conditioned reflexes. Studies were performed in dogs with chemotrodes implanted in the substantia nigra and the head of the caudate nucleus. 6-Hydroxydopamine was injected into the substantia nigra at a dose of 50 microg, and 10 microg of corticoliberin was injected into the caudate nucleus. Blood cortisol and catecholamine levels were determined. Analysis of the result showed that an interaction takes place in the neostriatum between the corticoliberin and dopaminergic systems, and that in conditions in which dopaminergic structures are excluded, the efficacy of corticoliberin in the performance of behavioral acts decreases by 30-40%, i.e., complete expression of its regulatory role of motor situational conditioned reflexes is lost.

Modification of the behavioral effects of corticoliberin by early post-natal administration of corticosteroid hormones.

Experiments on rats in which hydrocortisone was given in the early postnatal period were used to study the effects of intrastriatal microinjection of corticoliberin on behavior in an open field test. Bilateral microinjection of corticoliberin into the neostriatum led to a sharp reduction in orientational-investigative activity. Rats given hydrocortisone in the first days of life had elevated movement activity, and the anxiogenic effect of corticoliberin was absent in these animals.

Neuroendocrine mechanisms of the formation of adaptive behavior.

The behavioral and neuroendocrine responses of the body to external changes are determined by genetically determined programs of individual development, and are established during pre- and post-natal ontogenesis. These responses, however, can be changed by stress or administration of corticosteroid hormones in "critical periods" of the body's development. Mineralo- and glucocorticoid receptors mediate the "inhibition" of particular neuroendocrine or neuromediator systems, promoting behavioral modification.